The determination of estrone, estradiol-17beta, delta4-androstenedione, testosterone, progesterone, cortisol and corticosterone will be continued in the follicular fluid of the ovary at various stages of follicular development in pigs and in humans. The albumin which interferes in the examination of specific steroid binding proteins will be removed from the follicular fluid. The study of the binding of estradiol-17beta, testosterone and progesterone to the specific binding proteins of the follicular fluid from porcine and human ovaries will be continued. The physicochemical properties of the steroid binding proteins such as specificity, affinity, molecular size and other chemical properties will be investigated. The steroid binding protein(s) will be isolated by the affinity chromatography in a pure form and its physicochemical properties will be studied. The role of such binding protein component in the process of ovulation will be investigated. This could be achieved by its direct administration into follicle or a specific antibody will be produced in rabbit and then used to block the action of steroid binding component. If such component(s) is found physiologically active, it will be quantitated by radioimmunoassay. The site of production of steroid binding protein in the follicular fluid will be investigated. First, the properties of such steroid binding protein from follicular fluid will be compared to that of blood plasma. If found similar, the binding protein will be labeled and injected into pig or rat and its appearance in the follicle will be studied. Secondly, granulosa cells will be cultured and the possible synthesis and release of steroid binding component or other protein fraction that is contributed in the follicular fluid will be investigated. Specific steroid cytosol and nuclear receptor in the granulosa cells will be examined to correlate the fraction of steroids on the development of follicles and ovulation.